Integrated Systems for the Analysis of Circulating Multiple Myeloma Cells (CMMCs)

 描述（由申请人提供）：多发性骨髓瘤（MM）是一种浆细胞恶性肿瘤，约占恶性肿瘤的1%，占所有造血系统肿瘤的15%。根据美国癌症协会的数据，2014年美国估计有24，050例新病例被诊断出来，估计死亡人数为11，090人。意义不明的单克隆丙种球蛋白病（MGUS）每年进展为明显恶性肿瘤（最常见为MM）的风险为1%，在50岁以上的个体中发现约3%，患病率随年龄增加而增加。在没有明确的诊断测试来监测MGUS恶性进展的情况下，目前监测一系列因素，包括血清游离轻链比率的变化、升高的血清M蛋白水平和高骨髓浆细胞百分比。最近的研究已经证明，通过免疫荧光显微镜或流式细胞术测量的从骨髓活检中获得的循环多发性骨髓瘤细胞（CMMC）对于MM的诊断、预后和测量治疗反应是有用的。然而，获得骨髓活检对患者来说是痛苦的，这禁止了监测疾病进展、药物反应和/或复发所需的频繁测试。开发直接从外周血中选择和分析CMMC的新仪器和技术对于改善MM患者或其早期疾病阶段的管理至关重要。基于从I期类型研究中获得的大量初步数据，该II期项目专注于开发一种集成系统，该系统使用一次性射流盒和相关仪器外围设备，用于直接从全血中快速全自动处理CMMC，从而无需骨髓活检，从而允许更频繁地进行检测，特别是MGUS检测。该仪器将能够以>80%的回收率和>80%的纯度选择CMMC，释放所选择的细胞并对其进行电子计数，并将其准备用于FISH分析或免疫表型分析。该系统还可以卸载选定的CMMC进行分子分析，例如搜索KRAS突变。仪器的操作将自动化，样本至应答的总体处理时间约为1.5 h。该系统将由BioFluidica工作人员进行内部确认，并由查佩尔山的北卡罗来纳州大学的合作者进行外部确认。虽然MM将被用作仪器临床性能验证的案例研究，但通过编程正确选择抗体以搜索相关的血源性生物细胞，它可以很容易地用于不同的癌症相关疾病。

 DESCRIPTION (provided by applicant): Multiple myeloma (MM) is a plasma cell malignancy and comprises about 1% of malignant tumors and 15% of all hematopoietic neoplasms. According to the American Cancer Society, in 2014 an estimated 24,050 new cases were diagnosed in the U.S. with the estimated deaths being 11,090. Monoclonal gammopathy of undetermined significance (MGUS) is associated with a 1% per year risk of progression to overt malignancy, most commonly MM and is found in approximately 3% of individuals over 50 with the prevalence increasing with age. Without a clear diagnostic test to monitor MGUS malignancy progression, a range of factors are currently monitored including changes in serum free light chains ratio, elevated serum M protein level and high bone marrow plasma cell percentage . Recent studies have demonstrated that circulating multiple myeloma cells (CMMCs) secured from bone marrow biopsies measured by immuno-fluorescence microscopy or flow cytometry are useful for diagnosis, prognosis and measuring therapeutic response for MM. However, securing a bone marrow biopsy is painful to the patient prohibiting frequent testing required for monitoring disease progression, drug response and/or recurrence. Development of novel instrumentation and techniques for CMMC selection and analysis directly from peripheral blood is essential to improve the management of patients with MM or its early disease stages. Building on extensive preliminary data secured from a Phase I type study, this Phase II project is focused on the development of an integrated system that uses a disposable fluidic cartridge and the associated instrument peripherals for the rapid and fully automated processing of CMMCs directly from whole blood obviating the need for bone marrow biopsies and thus, allowing for more frequent testing especially those with MGUS. The instrument will be able to select CMMCs at >80% recovery and >80% purity, release the selected cells and electronically enumerate them and prepare them for FISH analysis or immunophenotyping. The system can also off-load the selected CMMCs for molecular profiling, such as searching for KRAS mutations. Operation of the instrument will be automated with an overall sample- to-answer processing time of ~1.5 h. The system will be subjected to both internal validation by BioFluidica staff as well as external validation by collaborators at the University of North Carolina at Chapel Hill. While MM will be used as a case study for clinical performance verification of the instrument, it can easily be utilized in different cancer-related diseases by programming in the correct selection antibody to search for the relevant blood-borne biological cells.